Spatiotemporal regulation of anther's tapetum degeneration paved the way for a reversible male sterility system in cotton

Male sterility is an important agronomical trait in self-pollinating plants for producing cost-effective F1 hybrids to harness the heterosis. Still, large-scale development and maintenance of male sterile lines and restoring fertility in F1 hybrids pose significant challenges in plant hybrid breeding. Cotton is a self-pollinating crop and exhibits strong hybrid vigor. However, there are currently few breeding methods to achieve cost-effective production of F1 hybrid cotton. Here, we utilized novel functions of the Arabidopsis autophagy-related BECLIN1/ATG6 and a mutant of E3 ubiquitin ligase COP1 (COP1^L105A) genes in developing rescuable male sterility in cotton. We have generated multiple male-sterile (MS) and restorer (RS) cotton lines expressing BECLIN1 and COP1^L105A, respectively. Cytological observation showed that post-meiotic tapetal expression of BECLIN1 delays tapetum developmental programmed cell death (dPCD) by affecting reactive oxygen species (ROS) balance-this delay in dPCD results in early microspore defects and later small-sized flowers with indehiscent anthers. Furthermore, the evaluation of F1 hybrids developed by crossing MS and RS lines showed that early tapetal COP1^L105A expression abolishes expression of BECLIN1 resulting in normal tapetum degeneration, pollen development, and fertility. In addition, the F1 hybrid developed with MS and RS cotton lines in transgenic glass-house and net-house conditions showed the rescued fertility comparable with control plants (WT). In terms of cotton fiber productivity, the COP1^L105A-expressing transgenic cotton lines outperformed the WT. The current work effectively demonstrates the wider applicability of the new F1 cotton production system.